Induction of Melanoma Cell-Selective Apoptosis Using Anti-HER2 Antibody-Conjugated Gold Nanoparticles
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Purpose: This study was conducted to verify the induction and mechanism of selective apoptosis in G361 melanoma cells usinganti-HER2 antibody-conjugated gold nanoparticles (GNP-HER2). Materials and Methods: Following GNP-HER2 treatment of G361 cells, cell cycle arrest and apoptosis were measured by WST-1assay, Hemacolor staining, Hoechst staining, immunofluorescence staining, fluorescence-activated cell sorting analysis, andWestern blotting. Results: G361 cells treated with GNP-HER2 showed condensation of nuclei, which is an apoptotic phenomenon, and translocationof apoptosis-inducing factor and cytochrome c from mitochondria into the nucleus and cytoplasm, respectively. Increases inBAX in cells undergoing apoptosis, activation of caspase-3 and -9, and fragmentation of poly (ADP-ribose) polymerase and DNAfragmentation factor 45 (inhibitor of caspase-activated DNase) were observed upon GNP-HER2 treatment. Following GNP-HER2treatment, an increase of cells in sub-G1 phase, which is a signal of cell apoptosis, was observed. This resulted in the down-regulationof cyclin A, cyclin D1, cyclin E, cdk2, cdk4, and cdc2 and the up-regulation of p21. Thus, GNP-HER2 treatment was confirmedto induce the cessation of cell cycle progression. Also, decreases in phospho-focal adhesion kinase and phospho-humanepidermal growth factor receptor, which activate cellular focal adhesion, and decreases in phospho-paxillin, which stimulates thedisassembly of filamentous actin, were observed. Reduced cell adhesion and disassembly of the intracellular structure indicatedcell deactivation. Conclusion: GNP-HER2 can selectively kill G361 melanoma cells without affecting normal cells. The mechanism of G361 celldeath upon treatment with GNP-HER2 was apoptosis accompanied by activation of caspases.
Abstract
Purpose: This study was conducted to verify the induction and mechanism of selective apoptosis in G361 melanoma cells usinganti-HER2 antibody-conjugated gold nanoparticles (GNP-HER2). Materials and Methods: Following GNP-HER2 treatment of G361 cells, cell cycle arrest and apoptosis were measured by WST-1assay, Hemacolor staining, Hoechst staining, immunofluorescence staining, fluorescence-activated cell sorting analysis, andWestern blotting. Results: G361 cells treated with GNP-HER2 showed condensation of nuclei, which is an apoptotic phenomenon, and translocationof apoptosis-inducing factor and cytochrome c from mitochondria into the nucleus and cytoplasm, respectively. Increases inBAX in cells undergoing apoptosis, activation of caspase-3 and -9, and fragmentation of poly (ADP-ribose) polymerase and DNAfragmentation factor 45 (inhibitor of caspase-activated DNase) were observed upon GNP-HER2 treatment. Following GNP-HER2treatment, an increase of cells in sub-G1 phase, which is a signal of cell apoptosis, was observed. This resulted in the down-regulationof cyclin A, cyclin D1, cyclin E, cdk2, cdk4, and cdc2 and the up-regulation of p21. Thus, GNP-HER2 treatment was confirmedto induce the cessation of cell cycle progression. Also, decreases in phospho-focal adhesion kinase and phospho-humanepidermal growth factor receptor, which activate cellular focal adhesion, and decreases in phospho-paxillin, which stimulates thedisassembly of filamentous actin, were observed. Reduced cell adhesion and disassembly of the intracellular structure indicatedcell deactivation. Conclusion: GNP-HER2 can selectively kill G361 melanoma cells without affecting normal cells. The mechanism of G361 celldeath upon treatment with GNP-HER2 was apoptosis accompanied by activation of caspases.
참고문헌 (32)
- Kim SH, "Microsomal PGE2 synthase-1 regulates melanoma cell survival and associates with melanoma disease progression", Pigment Cell Melanoma Res, 29, pp.297~308 (2016)
- Soengas MS, "Apoptosis and melanoma chemoresistance", Oncogene, 22, pp.3138~3151 (2003)
- Service RF, "Materials and biology. Nanotechnology takes aim at cancer", Science, 310, pp.1132~1134 (2005)
- Lam CW, "Pulmonary toxicity of single-wall carbon nanotubes in mice 7 and 90 days after intratracheal instillation", Toxicol Sci, 77, pp.126~134 (2004)
- Shi Kam NW, "Nanotube molecular transporters : internalization of carbon nanotube-protein conjugates into Mammalian cells", J Am Chem Soc, 126, pp.6850~6851 (2004)
- Magrez A, "Cellular toxicity of carbon-based nanomaterials", Nano Lett, 6, pp.1121~1125 (2006)
- Derfus AM, "Probing the cytotoxicity of semiconductor quantum dots", Nano Lett, 4, pp.11~18 (2004)
- Wu X, "Immunofluorescent labeling of cancer marker Her2 and other cellular targets with semiconductor quantum dots", Nat Biotechnol, 21, pp.41~46 (2003)
- Ahamed M, "ZnO nanorod-induced apoptosis in human alveolar adenocarcinoma cells via p53, survivin and bax", bcl-2 pathways : role of oxidative stress, 2011, pp.7, 904~913
- Arya G, "Enhanced antiproliferative activity of Herceptin(HER2)-conjugated gemcitabine-loaded chitosan nanoparticle in pancreatic cancer therapy", Nanomedicine, 7, pp.859~870 (2011)
- Connor EE, "Gold nanoparticles are taken up by human cells but do not cause acute cytotoxicity", Small, 1, pp.325~327 (2005)
- Thomas M, "Conjugation to gold nanoparticles enhances polyethylenimine’s transfer of plasmid DNA into mammalian cells", Proc Natl Acad Sci U S A, 100, pp.9138~9143 (2003)
- Rosenberg SJ, "Radical prostatectomy with adjuvant radioactive gold for prostatic cancer : a preliminary report", J Urol, 133, pp.225~227 (1985)
- Patra HK, "Cell selective response to gold nanoparticles", Nanomedicine, 3, pp.111~119 (2007)
- El-Sayed IH, "Surface plasmon resonance scattering and absorption of anti-EGFR antibody conjugated gold nanoparticles in cancer diagnostics : applications in oral cancer", Nano Lett, 5, pp.829~834 (2005)
- El-Sayed IH, "Selective laser photo-thermal therapy of epithelial carcinoma using anti-EGFR antibody conjugated gold nanoparticles", Cancer Lett, 239, pp.129~135 (2006)
- Bargmann CI, "The neu oncogene encodes an epidermal growth factor receptor-related protein", Nature, 319, pp.226~230 (1986)
- Rubin I, "The basic biology of HER2", Ann Oncol, 12(Suppl 1), pp.S3~8 (2001)
- Neve RM, "The role of overexpressed HER2 in transformation", Ann Oncol, 12(Suppl 1), pp.S9~13 (2001)
- Ullrich A, "Signal transduction by receptors with tyrosine kinase activity", Cell, 61, pp.203~212 (1990)
- Kämmerer U, "Expression of tumor markers on breast and ovarian cancer cell lines", Anticancer Res, 23, pp.1051~1055 (2003)
- Nakamura H, "Correlation between encoded protein overexpression and copy number of the HER2 gene with survival in non-small cell lung cancer", Int J Cancer, 103, pp.61~66 (2003)
- Ross JS, "The HER-2", neu oncogene in tumors of the gastrointestinal tract, 2001, pp.19, 554~568
- Oehler MK, "Molecular genetics and endometrial cancer", J Br Menopause Soc, 9, pp.27~31 (2003)
- Jacobson MD, "Programmed cell death in animal development", Cell, 88, pp.347~354 (1997)
- Nagata S, "Apoptosis by death factor", Cell, 88, pp.355~365 (1997)
- Alberts B, "Molecular biology of the cell" (2008), Garland Science
- Sabour Alaoui S, "TWEAK affects keratinocyte G2", M growth arrest and induces apoptosis through the translocation of the AIF protein to the nucleus, 2012, pp.7, e33609~
- Arias-Gonzílez I, "Cytotoxic effect of Kalanchoe flammea and induction of intrinsic mitochondrial apoptotic signaling in prostate cancer cells", J Ethnopharmacol, 222, pp.133~147 (2018)
- Fan Y, "Resveratrol induces autophagy-dependent apoptosis in HL-60 cells", BMC Cancer, 18, pp.581~ (2018)
- Luo T, "PARP-1 overexpression contributes to Cadmium-induced death in rat proximal tubular cells via parthanatos and the MAPK signalling pathway", Sci Rep, 7, pp.4331~ (2017)
- Errami Y, "ICAD deficiency in human colon cancer and predisposition to colon tumorigenesis : linkage to apoptosis resistance and genomic instability", PLoS One, 8, pp.e57871~ (2013)